Application of polygonatum sibiricum stem and leaf or extract thereof in preparation of feed or feed additive for improving animal growth performance and in preparation of medicine or health food with intestinal flora regulating function

Abstract

The invention provides application of polygonatum sibiricum stems and leaves or extract thereof in preparation of feed or feed additives for improving animal growth performance and in preparation of medicines or health-care foods with intestinal flora regulating functions, and belongs to the technical field of biological science. The invention finds that the rhizoma polygonati stems and leaves or the extract thereof can obviously improve the body weight of animals, reduce the feed conversion ratio and save feed, the effect is superior to that of rhizoma polygonati and rhizoma polygonati whiskers, and an unexpected technical effect is achieved. The invention also finds that the polygonatum sibiricum stems and leaves or the extract thereof can improve the microbial community of animal intestinal tracts, and has the functions of promoting the proliferation of probiotics and inhibiting the growth of harmful bacteria. The polygonatum sibiricum stems and leaves and the extract thereof are used for preparing feed or feed additives for improving the growth performance of animals, and medicines or health-care foods with the function of regulating intestinal flora, and the polygonatum sibiricum stems and leaves and the extract thereof have wide market application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of biological science and technology, and specifically relates to the use of polygonatum stems and leaves or extracts thereof in preparing feed or feed additives for improving animal growth performance, and in preparing medicines or health foods with the function of regulating intestinal flora. Background Art

[0002] The Chinese medicinal herb Polygonatum sibiricum is the dried rhizome of Polygonatum kingianum (Coll. et Hemsl), Polygonatum sibiricum (Red.), or Polygonatum cyrtonema Hua (Hua), all members of the Liliaceae family. Depending on its shape, it is commonly known as "big polygonatum," "chicken-headed polygonatum," or "ginger-shaped polygonatum." It boasts benefits such as tonifying qi, strengthening the spleen, moistening the lungs, and benefiting the kidneys. In recent years, its use in poultry farming has gained increasing attention. The rhizome of Polygonatum sibiricum is the medicinal part of the pharmacopoeia, while the aboveground stems and leaves have been less utilized and long considered non-medicinal or agricultural waste, resulting in low resource utilization.

[0003] In recent years, with increasing emphasis on the comprehensive development of medicinal plant resources, research has revealed that the stems and leaves of Polygonatum sibiricum contain active ingredients such as flavonoids, polysaccharides, saponins, alkaloids, and phenols. However, due to traditional usage practices and insufficient research, the development and utilization of Polygonatum sibiricum stems and leaves remains in its early stages, and a systematic functional product system has yet to be established. Therefore, exploring the functional activities of Polygonatum sibiricum stems and leaves is of great significance. On the one hand, this can fully utilize Polygonatum sibiricum resources and reduce waste; on the other hand, it can facilitate the development of more new health foods or pharmaceuticals with specialized functions, meet consumer demand for health products, and promote the sustainable development of the Polygonatum sibiricum industry. Summary of the Invention

[0004] The purpose of the present invention is to provide the use of polygonatum stems and leaves or extracts thereof in preparing feed or feed additives for improving animal growth performance and in preparing medicines or health foods with the function of regulating intestinal flora.

[0005] The invention provides use of polygonatum stems and leaves or extracts thereof in preparing feed or feed additives for improving animal growth performance.

[0006] The "stems and leaves of Polygonatum sibiricum" of the present invention refer to the above-ground stems and leaves of the Chinese medicinal material Polygonatum sibiricum.

[0007] The present invention also provides the use of the stems and leaves of polygonatum or the extract thereof in preparing medicines, health foods, feeds or feed additives with the function of regulating intestinal flora.

[0008] Furthermore, the medicine, feed or feed additive is a medicine, feed or feed additive for preventing and / or treating obesity, diabetes or diarrhea;

[0009] The health food is a health food that helps to enhance immunity, control body fat, and maintain healthy blood sugar levels.

[0010] Furthermore, the medicine, health food, feed or feed additive is a substance that increases the richness of intestinal flora; and / or, the medicine, health food, feed or feed additive is a substance that promotes the proliferation of probiotics and inhibits the growth of harmful bacteria.

[0011] Furthermore, the intestinal flora belongs to the phylum Verrucomicrobia;

[0012] The probiotics are Lactobacillus, Akkermansia, Allobaculum, Lachnospiraceae_Clostridium and Flexispira;

[0013] The harmful bacteria are Helicobacter and Prevotella.

[0014] Furthermore, each ton of feed or feed additive contains 1-8 kg of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 1-8 kg of polygonatum stems and leaves, preferably contains 5 kg of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 5 kg of polygonatum stems and leaves.

[0015] Furthermore, the unit preparation of the medicine or health food contains 18.9-57.4g of Polygonatum sibiricum stems and leaves or a Polygonatum sibiricum stem and leaf extract prepared from 18.9-57.4g of Polygonatum sibiricum stems and leaves, preferably 38.5g of Polygonatum sibiricum stems and leaves or a Polygonatum sibiricum stem and leaf extract prepared from 38.5g of Polygonatum sibiricum stems and leaves.

[0016] Furthermore, the dosage form of the medicine or health food is one or more of capsules, tablets, oral liquids, and granules;

[0017] The feed or feed additive is in the form of one or more of powder, granules, and premixes.

[0018] Furthermore, the medicine, health food, feed or feed additive also includes one or more of the following excipients: filler, binder, disintegrant, lubricant, flavoring agent, colorant.

[0019] Furthermore, the extract of polygonatum stem and leaf is a water extract of polygonatum stem and leaf, an alcohol extract of polygonatum stem and leaf, or a mixture of a water extract of polygonatum stem and leaf and a polygonatum stem and leaf extract, preferably a water extract of polygonatum stem and leaf.

[0020] The present invention also provides a feed or feed additive, wherein each ton of the feed or feed additive contains 1-8 kg of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 1-8 kg of polygonatum stems and leaves.

[0021] Furthermore, each ton of feed or feed additive contains 5 kg of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 5 kg of polygonatum stems and leaves.

[0022] The present invention also provides a medicine or health food, wherein a unit preparation of the medicine or health food contains 18.9-57.4 g of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 18.9-57.4 g of polygonatum stems and leaves.

[0023] Furthermore, a unit preparation of the medicine or health food contains 38.5 g of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 38.5 g of polygonatum stems and leaves.

[0024] The conversion method for the content of Polygonatum sibiricum stem and leaf or its extract in a unit dose of medicine or health food in the present invention is as follows: according to the preparation method of Example 2, the dosage of the Polygonatum sibiricum stem and leaf extract for mice is 5g / kg in terms of the raw material drug. According to the "Pharmacological Test Methodology" edited by Xu Shuyun, the dosage for mice = 9.1 × the dosage for humans (body weight 70kg). Therefore, the dosage for humans (body weight 70kg) converted from the dosage group of the Polygonatum sibiricum stem and leaf extract is 0.55g / kg. Therefore, the content of Polygonatum sibiricum stem and leaf or its extract in a unit dose of medicine or health food is 0.55g / kg × 70kg = 38.5g.

[0025] The unit dosage form of the present invention is calculated based on an average adult weight of 70 kg.

[0026] The present invention has achieved the following beneficial effects:

[0027] (1) The present invention found that the stems and leaves of Polygonatum sibiricum can significantly increase the weight gain of poultry, and the effect is better than that of Polygonatum sibiricum and Polygonatum sibiricum, achieving unexpected technical effects;

[0028] (2) The stems and leaves of Polygonatum sibiricum can reduce the feed-to-meat ratio and save feed, which has promoted the development and industrial application of the stems and leaves of Polygonatum sibiricum in feeds that improve the growth performance of poultry and has broad market prospects;

[0029] (3) The present invention further optimized the process conditions for extracting the Polygonatum sibiricum stem and leaves, and screened out the preferred process conditions shown in Example 2;

[0030] (4) Polygonatum sibiricum stems and leaves or their extracts can significantly increase the body weight of normal mice and cyclophosphamide-immunosuppressed mice;

[0031] (5) Polygonatum sibiricum stems and leaves or their extracts have an excellent regulatory effect on the distribution of immune cell numbers caused by cyclophosphamide immunosuppression, and the best regulatory effect is obtained with a medium dose.

[0032] (6) Polygonatum sibiricum stems and leaves or their extracts can significantly increase the serum IFN-γ, IgM, and IgG levels in mice with cyclophosphamide immunosuppression model, with the medium-dose group showing the best effect;

[0033] (7) The stems and leaves of Polygonatum sibiricum or its extract can regulate the intestinal flora function of cyclophosphamide immunosuppressed mice, significantly increase the relative abundance of Verrucomicrobia to normal levels, alleviate the intestinal flora imbalance caused by immunosuppression, promote the proliferation of probiotics (Lactobacillus, Akkermansia, Allobaculum, Lachnospiraceae_Clostridium, Flexispira), and inhibit the growth of harmful bacteria (Helicobacter, Prevotella), and the effect is better than that of Astragalus extract.

[0034] Obviously, based on the above contents of the present invention, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0035] The following is a further detailed description of the present invention through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-mentioned content of the present invention fall within the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 Analysis of the α-diversity of intestinal flora: (A) Chao1, (B) Observed_species, (C) Faith's PD, (D) Shannon and (E) Simpson (Note: the horizontal axis represents different groups; the vertical axis represents the α-diversity index).

[0037] Figure 2 Figure β-diversity analysis (PCA) of intestinal flora: (A) The second principal component is PC2; (B) The second principal component is PC3 (Note: the horizontal axis represents the first principal component, the vertical axis represents the second principal component, and the percentage represents the contribution to the sample difference).

[0038] Figure 3 (A) Species composition and difference analysis at the phylum level (top 20) (Note: (left) Different colors represent species with different metabolisms, corresponding to the legend on the right; the horizontal axis represents different groups, and the vertical axis represents the relative abundance of different species; (right) the horizontal axis represents different groups, and the vertical axis represents different species. The depth of color is related to the abundance of the species, and the darker the color, the higher the abundance). (B) Species composition and difference analysis at the phylum level.

[0039] Figure 4 (A) Species composition and difference analysis at the genus level (top 20) (Note: (left) Different colors represent species with different metabolisms, corresponding to the legend on the right; the horizontal axis represents different groups, and the vertical axis represents the relative abundance of different species; (right) the horizontal axis represents different groups, and the vertical axis represents different species. The depth of color is related to the abundance of the species, and the darker the color, the higher the abundance). (B) Species composition and difference analysis at the genus level. DETAILED DESCRIPTION

[0040] The raw materials and equipment used in the present invention are all known products and are obtained by purchasing commercially available products.

[0041] Example 1: Effect of Polygonatum sibiricum Stems and Leaves on Improving Poultry Growth Performance

[0042] 1. Experimental Materials

[0043] 1.1 Experimental Animals

[0044] 240 laying hens (Hylan Grey), 50 days old, were purchased from a large poultry enterprise in Sichuan.

[0045] 1.2 Medicinal Materials and Reagents

[0046] Polygonatum multiflorum (the main medicinal part is its root), grind into 80 meshes with a grinder and set aside;

[0047] The stems and leaves of Polygonatum cyrtonema are crushed into 80 meshes by a grinder and set aside;

[0048] The roots of Polygonatum sibiricum are crushed into 80 meshes by a grinder and set aside.

[0049] 2. Experimental methods

[0050] 2.1. Trial Grouping and Dosing

[0051] A total of 240 50-day-old Hy-Line Grey chickens were randomly assigned to three experimental groups (Groups D, E, and F), two drug control groups (Groups B and C), and one blank control group (Group A), with 40 chickens in each group. Four parallel groups were included within each group. The experimental design is shown in Table 1. The experimental groups maintained the same dietary composition, nutritional level, environmental conditions, and daily care requirements as the control group. The entire trial lasted 15 days.

[0052] Table 1 Experimental design and dosage

[0053] Grouping Dosage Quantity (pieces) Blank control group (Group A) No medication 40 Polygonatum sibiricum group (Group B) 3kg / ton feed 40 Polygonatum sibiricum group (Group C) 3kg / ton feed 40 High-dose group of Polygonatum sibiricum stems and leaves (Group D) 5kg / ton feed 40 Middle dose group of Polygonatum sibiricum stems and leaves (Group E) 3kg / ton feed 40 Low-dose group of Polygonatum sibiricum stems and leaves (Group F) 1kg / ton feed 40

[0054] 2.2 Observation indicators

[0055] The weight of each experimental group and the control group was recorded before administration. They were weighed 15 days after administration, and the total feed intake was recorded to calculate the feed-to-meat ratio. The feed-to-meat ratio was calculated as follows:

[0056]

[0057] 3. Experimental results

[0058] Table 2 Effects of Polygonatum sibiricum stems and leaves on chicken production performance

[0059]

[0060] After 15 days of feeding with the three medicinal herbs, the experimental results (Table 2) showed that the high, medium, and low dose groups of Polygonatum sibiricum stems and leaves all promoted weight gain in chickens, with the high dose group showing the best effect. From an economic perspective, the medium dose group is recommended. The results showed that Polygonatum sibiricum and Polygonatum sibiricum whiskers could also promote weight gain in chickens, but the Polygonatum sibiricum stem and leaf group was more effective than the Polygonatum sibiricum and Polygonatum sibiricum whiskers groups. The high, medium, and low dose groups of Polygonatum sibiricum stem and leaf only gained an average of 273.08g, 269.63g, and 248.92g, respectively. The dosage of the medium dose group of Polygonatum sibiricum stem and leaf, Polygonatum sibiricum group, and Polygonatum sibiricum whiskers group was 3kg / ton of feed, but the weight gain effect of the medium dose group of Polygonatum sibiricum stem and leaf was 20.05g and 27.75g higher than the Polygonatum sibiricum medicinal herb group and Polygonatum sibiricum whiskers group, respectively, and 41.09g higher than the control group. The feed-to-meat ratio was also significantly lower than that of the Polygonatum sibiricum medicinal material group and the Polygonatum sibiricum whisker group, indicating that the stems and leaves of Polygonatum sibiricum can significantly increase the weight gain of chickens, reduce the feed-to-meat ratio, save feed, and the effect is better than that of the Polygonatum sibiricum and Polygonatum sibiricum whisker groups, achieving unexpected technical results.

[0061] Example 2: Effects of Polygonatum sibiricum Stem and Leaf Extract on Immune Function and Intestinal Flora in Cyclophosphamide-Immunosuppressed Mice

[0062] 1. Experimental Materials

[0063] 1.1 Experimental Animals

[0064] 84 SPF-grade ICR mice, half male and half female, weighing 16-18 g, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., with the experimental animal production license number SCXK (Beijing) 2019-0010.

[0065] 1.2 Reagents and drugs

[0066] Cyclophosphamide (CTX, Shanghai Yuanye Biotechnology Co., Ltd., batch number F13IS206786);

[0067] Mouse IgM (catalog number: RX202732M), IgG (catalog number: RX202736M), and interferon-γ (catalog number: RX203097M) enzyme-linked immunosorbent assay kits were purchased from Quanzhou Ruixin Biotechnology Co., Ltd. (China); Elab Red 780 Anti-Mouse CD3 Antibody [17A2] (Cat. No.: E-AB-F1013S), PE / Cyanine5.5 Anti-Mouse CD4 Antibody [GK1.5] (Cat. No.: E-AB-F1097I), and PE / Cyanine7 Anti-Mouse CD8a Antibody [53-6.7] (Cat. No.: E-AB-F1104H) were purchased from Elabscience (China); BD Pharmingen TM PE Rat Anti-Mouse F4 / 80 (Cat. No. 565410), BD Pharmingen TM APC Rat Anti-Mouse CD49b (Cat. No. 560628) and BD Pharmingen TM FITC Rat Anti-Mouse CD19 (Cat. No. 557398) was purchased from BD Pharmingen TM Company (USA).

[0068] Astragalus extract: Take 1 kg of Astragalus root, add 15 kg of water, decoct twice at 90-100°C, each time for 1 hour, combine the decoctions, and vacuum dry at 60°C to obtain the Astragalus extract, which is set aside;

[0069] Polygonatum sibiricum stem and leaf extract: Take 1 kg of Polygonatum sibiricum stem and leaf, add 20 kg of water, decoct twice at 90-100°C, each time for 1 hour, combine the decoctions, and vacuum dry at 60°C to obtain the Polygonatum sibiricum stem and leaf extract, and set aside.

[0070] 1.3 Instruments

[0071] ZE5 Flow Cytometer

[0072] 2. Experimental methods

[0073] 2.1 Modeling and drug administration

[0074] The mice were adaptively fed for 3 days and then randomly divided into control group (G1), cyclophosphamide model group (G2), astragalus extract group (G3), low-dose polygonatum stem and leaf extract group (G4), medium-dose polygonatum stem and leaf extract group (G5), high-dose polygonatum stem and leaf extract group (G6), and polygonatum stem and leaf extract control group (G8, no modeling, only drug administration), with 12 mice in each group.

[0075] The Astragalus extract group (G3) was orally administered with 5 g / kg (calculated as the original medicinal material, i.e., astragalus medicinal material) of Astragalus extract;

[0076] The low (G4), medium (G5), and high (G6) dose groups of Polygonatum sibiricum stem and leaf extract and the control group (G8) of Polygonatum sibiricum stem and leaf extract were gavage-administered at 2.5 g / kg, 5.0 g / kg, 7.5 g / kg, and 5.0 g / kg (calculated as the original medicinal material, i.e., calculated as the Polygonatum sibiricum stem and leaf) body weight, respectively;

[0077] The control group (G1) and cyclophosphamide model group (G2) were given an equal volume of normal saline by gavage;

[0078] Each group was continuously administered with drug or normal saline for 15 days. On the 8th to 10th day after administration, mice in groups G2, G3, G4, G5, and G6 were intraperitoneally injected with 80 mg / kg cyclophosphamide to establish models. After the experiment, samples from each group were collected for testing of various indicators.

[0079] 2.2. Measurement of mouse body weight and food intake

[0080] During the administration period, the body weight and water intake of mice in each group were recorded every day, and the changes in body weight and water intake were statistically analyzed after 14 days.

[0081] 2.3. Detection of immune cell levels in blood samples

[0082] At the end of the experiment, blood was collected from the orbital cavity of each group. 100 μl of blood was placed in a flow cytometer. 1 μg of each CD4+, CD8+, F4 / 80, CD19, and NK1.1 antibody was added, mixed, and stained at 4°C in the dark for 30 minutes. 2 ml of lysis working solution was added, and the cells were incubated at room temperature for 10 minutes. 2 ml of PBS was added, and the cells were centrifuged at 300 g for 5 minutes. The supernatant was discarded, and 2 ml of PBS was added. The cells were resuspended in 400 μl of PBS and analyzed using a ZE5 flow cytometer.

[0083] 2.4. Serum IFN-γ, IgM, and IgG Levels

[0084] Blood samples from each group were centrifuged at 3000 rpm for 10 min, and serum was separated. Serum IFN-γ, IgM, and IgG levels were determined using the double antibody sandwich method according to the ELISA kit instructions.

[0085] 2.5 Analysis of intestinal flora diversity

[0086] After the administration, the cecal contents of 6 male mice in each group were collected, and the samples were placed in sterile EP tubes and stored at -80 ° C. All samples were sent to Shenzhen Micro-Technology Group Co., Ltd. for sequencing. The genomic DNA of the samples was extracted using CTAB, and the V3+V4 variable regions were PCR amplified using primers 341F (5'-CCTAYGGGRBGCASCAG-3') and 806R (5'-GGACTACNNGGGTATCTAAT-3') and sequenced using the NovaSeq 6000PE250. Data processing and analysis were then performed on the WekemoBioincloud (https: / / www.bioincloud.tech) online cloud platform, and the comparison database was the Greengenes2 database (16S).

[0087] 2.6 Data Statistics and Analysis

[0088] All data were analyzed using GraphPad Prism software, and the results are presented as mean ± SD. Statistical inference was performed using one-way analysis of variance, followed by the Turkey test. Significant differences are indicated with *p < 0.05, and extremely significant differences are indicated with **p < 0.01.

[0089] 3. Experimental results and analysis

[0090] 3.1 Effects of mouse body weight and food intake

[0091] After 14 days of feeding, the weight of mice in the model group decreased by 0.56g, indicating that the model was successfully established; the weight of mice in the drug-administered model group increased to a certain extent, and the difference was significant compared with the model group, with the medium and high dose groups having better effects. The weight gain of mice in the normal group of Polygonatum sibiricum stems and leaves was significantly different from that in the control group without drug administration, indicating that Polygonatum sibiricum stems and leaves can significantly increase the weight of normal mice and immune model mice. The results are shown in Table 3. Figure 1 (A~E).

[0092] Table 3 Effects of Polygonatum sibiricum stems and leaves on body weight and food intake of immunosuppressive model mice

[0093]

[0094] 3.2. Detection of immune cell levels in blood samples

[0095] The results showed that compared with the normal group, the number of immune cells, including CD3+, CD8+, CD49b+, and CD19+, decreased in the model group (p ≤ 0.01). There was no significant difference in the number of CD4+ cells (p ≥ 0.05). F4 / 80+ cells also decreased, indicating successful modeling. High, medium, and low doses of Polygonatum sibiricum stems and leaves significantly modulated the number of immune cells caused by immunosuppression, with the medium dose showing the greatest effect. There was no significant difference in the number of CD3+, CD4+, and CD8+ cells compared with the normal group. The CD4+ / CD8+ ratio is an important indicator of whether the body's immunity is unbalanced. It is generally believed that the normal ratio is about 2. This experiment showed that the CD4+ / CD8+ ratio of the normal group was 2.15, indicating that the mouse's immunity was in a normal state, and the ratio of the model group was 2.72, indicating that it was in a serious imbalance state. The CD4+ / CD8+ ratios of the high, medium and low dose groups of Polygonatum sibiricum stems and leaves were all lower than those of the model group, indicating that Polygonatum sibiricum stems and leaves can significantly regulate the body's immune imbalance. The medium dose group had the best effect, and there was no significant difference compared with the normal group, indicating that it has returned to normal.

[0096] Table 4 Effects of Polygonatum sibiricum stems and leaves on the distribution of immune cells in immunosuppressive model mice (%)

[0097]

[0098] Note: Compared with the normal group: *p<0.05, **p<0.01; compared with the medium-dose group: #p<0.05, ##p<0.01.

[0099] 3.3. Serum IFN-γ, IgM, and IgG Levels

[0100] The test results showed that the stems and leaves of Polygonatum sibiricum could significantly increase the serum IFN-γ, IgM, and IgG levels in immunosuppressive model mice, with the medium-dose group showing the best effect. The normal group and the normal group of Polygonatum sibiricum stem and leaf extracts were not treated with cyclophosphamide to establish the model. The test results showed that feeding the Polygonatum sibiricum stem and leaf extract to normal mice also significantly increased the serum IFN-γ, IgM, and IgG levels. The results are shown in Table 5.

[0101] Table 5 Effects of Polygonatum sibiricum stems and leaves on the distribution of immune cells in immunosuppressive model mice (%)

[0102]

[0103]

[0104] Note: Compared with the normal group: *p<0.05, **p<0.01; compared with the medium-dose group: #p<0.05, ##p<0.01.

[0105] 3.4 Effects of Polygonatum sibiricum Stem and Leaf Extract on Intestinal Microflora in Immunosuppressed Mice

[0106] 3.4.1. Microbial diversity analysis

[0107] The α-diversity index can evaluate the diversity and richness of the intestinal flora. This study used five commonly used indices, Observed species, Chao1, Shannon, Simpson, and Faith's PD, to measure diversity and richness. The Wilcoxon rank sum test was used to analyze the sample data of each of the above indices to screen out the α-diversity indices with significant differences among the sample groups. Figure 1 As shown in (A-E), there was no significant difference in the α-diversity index among the groups (P>0.05), indicating that short-term use of Polygonatum sibiricum stem and leaf extract had no significant effect on the species richness and uniformity of the intestinal flora in mice.

[0108] β-diversity is the comparison of microbial community composition between different samples (groups). Figure 2 As shown in Figures (A-B), the intestinal flora of the cyclophosphamide model group differed somewhat from that of the normal group, indicating that cyclophosphamide can cause changes in the species composition of the mouse intestinal flora. The composition of the intestinal flora of mice treated with Astragalus and Polygonatum sibiricum stem and leaf also differed from that of the normal group, with the Polygonatum sibiricum stem and leaf extract treatment being closer to that of the normal group. Furthermore, the microbial community structure of the groups treated with different Astragalus and Polygonatum sibiricum stem and leaf extracts also differed, suggesting that Polygonatum sibiricum stem and leaf have a certain regulatory effect on the intestinal flora.

[0109] 3.4.2. Microbial community structure analysis

[0110] like Figure 3As shown in (A-B), at the phylum level, Bacteroidetes and Firmicutes were the primary components of the mouse intestinal microbiota, accounting for over 90% of the total microbial population, followed by Proteobacteria and Verrucomicrobia. The proportions of Firmicutes and Bacteroidetes were similar in the normal control group. However, Firmicutes decreased after CTX induction, but increased to varying degrees after feeding Polygonatum sibiricum stem and leaf extract. Bacteroidetes also increased to varying degrees in both the CTX model group and the Polygonatum sibiricum stem and leaf extract group. Compared with the normal control group, the relative abundance of Proteobacteria increased in the cyclophosphamide model group, while the relative abundance of Verrucomicrobia decreased sharply. This suggests that immunosuppression can disrupt the original bacterial structure of mice, leading to a large increase in harmful bacteria and causing intestinal flora disorder in mice. After intervention with Polygonatum sibiricum stem and leaf extract, the relative abundance of Proteobacteria decreased significantly, even lower than that of the normal group; the relative abundance of Verrucomicrobia increased significantly to normal levels, alleviating the intestinal flora imbalance caused by immunosuppression. At the same time, compared with the normal group, the relative abundance of Proteobacteria in the Polygonatum sibiricum stem and leaf extract control group was also significantly reduced.

[0111] like Figure 4 As shown in (A-B), at the genus level, compared with the normal group, except for the low-dose group of Polygonatum sibiricum stem and leaf extract, the relative abundance of Bacteroides in the other CTX modeling groups increased to varying degrees. Moreover, after CTX modeling, the relative abundance of beneficial bacterial genera such as Lactobacillus, Akkermansia, Allobaculum, Lachnospiraceae_Clostridium, and Flexispira decreased sharply, while they increased to varying degrees after intervention with the extracts. Among them, the promoting effect of Polygonatum sibiricum stem and leaf extract was better than that of Astragalus extract. At the same time, compared with the normal group, the relative abundance of harmful bacterial genera such as Helicobacter and Prevotella in the model group increased significantly, while the relative abundance decreased after intervention with the extracts, even lower than that of the normal group. Among them, Polygonatum sibiricum stem and leaf extract was still better than Astragalus extract. Therefore, Polygonatum sibiricum stem and leaf extract has the function of promoting the proliferation of probiotics and inhibiting the growth of harmful bacteria, and its effect is better than that of Astragalus extract.

[0112] The above experimental results show that the stems and leaves of Polygonatum sibiricum can significantly increase the body weight of normal mice and immune model mice; the high, medium and low dose groups of Polygonatum sibiricum stems and leaves have a good regulatory effect on the number distribution of immune cells caused by immunosuppression, and the medium dose has the best regulatory effect overall; the stems and leaves of Polygonatum sibiricum can significantly increase the serum IFN-γ, IgM and IgG levels of immunosuppressive model mice, and the medium dose group has the best effect; the extract of Polygonatum sibiricum stems and leaves can regulate the intestinal flora function of cyclophosphamide immunosuppressed mice, and the relative abundance of Verrucomicrobia (Verrucomicrobia) is significantly increased to normal levels, alleviating the intestinal flora imbalance caused by immunosuppression, and has the function of promoting the proliferation of probiotics (Lactobacillus, Akkermansia, Allobaculum, Lachnospiraceae_Clostridium, Flexispira) and inhibiting the growth of harmful bacteria (Helicobacter, Prevotella), and the effect is better than that of Astragalus extract.

[0113] In summary, the present invention provides the use of the stems and leaves of Polygonatum sibiricum or their extracts in the preparation of feed or feed additives for improving animal growth performance, and in the preparation of medicines or health foods with the function of regulating intestinal flora. The present invention found that the stems and leaves of Polygonatum sibiricum or their extracts can significantly increase animal body weight, reduce feed-to-meat ratio, save feed, and the effect is better than that of Polygonatum sibiricum and Polygonatum sibiricum, and an unexpected technical effect has been achieved. The present invention also found that the stems and leaves of Polygonatum sibiricum or their extracts can improve the microbial community in the intestines of animals, have the function of promoting the proliferation of probiotics and inhibiting the growth of harmful bacteria. The use of the stems and leaves of Polygonatum sibiricum and their extracts to prepare feed or feed additives for improving animal growth performance, as well as medicines or health foods with the function of regulating intestinal flora, has broad market application prospects.

Claims

1. Use of the stems and leaves of Polygonatum sibiricum or their extracts in preparing feed or feed additives for improving animal growth performance.

2. Use of the stems and leaves of Polygonatum sibiricum or their extracts in the preparation of medicines, health foods, feeds or feed additives with the function of regulating intestinal flora.

3. The use according to claim 2, characterized in that The medicine, feed or feed additive is a medicine, feed or feed additive for preventing and / or treating obesity, diabetes or diarrhea; The health food is a health food that helps to enhance immunity, control body fat, and maintain healthy blood sugar levels.

4. The use according to claim 3, characterized in that The medicine, health food, feed or feed additive is a substance that increases the richness of intestinal flora; and / or, the medicine, health food, feed or feed additive is a substance that promotes the proliferation of probiotics and inhibits the growth of harmful bacteria.

5. The use according to claim 4, characterized in that The intestinal flora belongs to the phylum Verrucomicrobia; The probiotics are Lactobacillus, Akkermansia, Allobaculum, Lachnospiraceae_Clostridium and Flexispira; The harmful bacteria are Helicobacter and Prevotella.

6. The use according to any one of claims 1 to 5, characterized in that The polygonatum stem and leaf extract is a polygonatum stem and leaf water extract, a polygonatum stem and leaf alcohol extract, or a mixture of a polygonatum stem and leaf water extract and a polygonatum stem and leaf extract, preferably a polygonatum stem and leaf water extract.

7. A feed or feed additive, characterized in that Each ton of feed or feed additive contains 1-8 kg of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 1-8 kg of polygonatum stems and leaves.

8. The feed or feed additive according to claim 7, characterized in that Each ton of feed or feed additive contains 5kg of polygonatum stems and leaves or polygonatum stem and leaf extract prepared from 5kg of polygonatum stems and leaves.

9. A medicine or health food, characterized in that: A unit preparation of the medicine or health food contains 18.9-57.4g of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 18.9-57.4g of polygonatum stems and leaves.

10. The medicine or health food according to claim 9, characterized in that A unit preparation of the medicine or health food contains 38.5g of polygonatum stems and leaves or a polygonatum stem and leaf extract prepared from 38.5g of polygonatum stems and leaves.

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